人抗Aβ_(42)自身抗体的纯化及鉴定

Purification and identification of anti-Aβ_(42) autoantibody in healthy human serum

目的纯化并鉴定正常人血清中抗Aβ42自身抗体。方法采集正常人血清标本,测定抗体含量,用饱和硫酸铵沉淀血清得到IgG粗品,用CNBr Sephrose 4B层析柱对粗品IgG进一步纯化,采用PAGE电泳和Western blot方法鉴定IgG纯度和免疫活性。结果用CNBr活化Sephrose 4B柱可纯化出较高纯度的抗Aβ42自身抗体,纯度可达到90%。结论 CNBr活化Sephrose 4B层析柱可有效地从人血清中纯化出较高纯度的抗Aβ42自身抗体,便于对自身抗体进一步研究,为AD的免疫治疗提供更多的信息。

Objective To purify and identify the anti- Aβ42 autoantibody in normal human serum. Methods The normal human serum was collected to determine the concentration of the antibody. The serum was precipitated with saturated ammonium sulfate to get crude serum IgG. The IgG was further isolated and purified by affinity chromatography on Aβ42 immuno-affinity column. Then PAGE and Western blot was applied to determine the purity and immune-activity of IgG. Results The Sephrose 4B affinity column activated with CNBr can produce a highly purified anti- Aβ42 autoantibody and the purity rate is high up to 90%. Conclusion The Sephrose 4B affinity column activated with CNBr can effectively produce a highly purified anti- Aβ42 autoantibody from human serum. It will facilitate further study on autoantibody and provide more information for immunization treatment of AD.